大鲵皮胶原蛋白肽的抗原反应特性研究

Antigen response properties of collagen peptides from Andrias davidianus skin

中文关键词:  大鲵  胶原蛋白肽  酶联免疫吸附反应  抗原性

英文关键词:Andrias davidianus  collagen peptide  enzyme linked immunosorbent assay  antigenicity

中文摘要:

      目的 探讨酶法提取大鲵(Andrias davidianus)皮胶原蛋白肽的抗原性, 为进一步研究大鲵皮胶原蛋白肽的生物相容性提供依据。方法 采用酶法提取获得大鲵皮胶原蛋白肽, 并对得到的胶原蛋白肽进行基质辅助激光解吸电离飞行时间质谱法(matrix assisted laser desorption/ionization time-of-flight mass spectrometry, MALDI-TOF-MS)分析, 确定其分子量分布范围。将得到的胶原蛋白肽与弗氏佐剂混合乳化后经免疫程序免疫ICR (Institute of Cancer Research)小鼠, 采用抗体酶联免疫试剂盒检测小鼠血清中IgG、IgA、IgM及IgE含量。结果 酶解后的胶原蛋白肽分子量均小于10 kDa, 多数肽分子量集中在2000~5000 m/z, 检测血清中IgA、IgG、IgM和IgE 4种免疫球蛋白的质量浓度范围分别是71.63~85.90 ?g/mL、705.19~857.04 ng/mL、47.04~57.46 ng/mL与224.54~261.0 ?g/mL。结论 采用碱性蛋白酶酶解获得的大鲵皮胶原蛋白肽对ICR小鼠呈现出较弱的抗原性。

英文摘要:

      Objective To investigate the antigenicity of collagen peptide extracted from Andrias davidianus skin by enzymatic method, and provide data for further studying on its biocompatibility. Methods The collagen peptide from Andrias davidianus skin was extracted by alkaline proteinase, and its molecular weight distribution range was determined by matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The obtained collagen peptides were mixed and emulsified with Freund’s adjuvant, and ICR (Institute of Cancer Research) mice were immunized with immune program, IgG, IgA, IgM and IgE content in mouse serum were detected by antibody enzyme linked immunosorbent assay kits. Results The molecular weights of the collagen peptide after enzymolysis were less than 10 kDa, and the molecular weights of most peptides were concentrated in 2000?5000 m/z, and the mass concentration ranges of IgA, IgG, IgM and IgE in the detected serum were 71.63?85.90 ?g/mL, 705.19?857.04 ng/mL, 47.04?57.46 ng/mL and 224.54?261.0 ?g/mL, respectively. Conclusion The Andrias davidianus skin collagen peptide shows weak antigenicity to ICR mice.